1. Technical Field
The present invention relates to a chromatographic quantitative assay device for assaying an analyte utilizing antigen-antibody reaction and, more particularly, a chromatographic quantitative assay device including a liquid-impermeable sheet member, and a manufacturing method thereof.
2. Background Art
Conventionally, an assay method by immunochromatography utilizing antigen-antibody reaction is universally taken as an implementation method for chemical or clinical examination of a liquid sample, such as water analysis or urinalysis. Generally, chromatography refers to a method for separating mixtures in accordance with their components, and a specimen used for chromatography comprises a plurality of portions as follows: a portion for adding a liquid sample, a portion for holding a marker reagent which can migrate by infiltration of the liquid sample, a portion that performs binding reaction with the marker reagent containing a substance capable of being bound specifically to a flowing-down analyte, and a portion for absorbing a flowing-down sample. According to this assay method of a chromatographic assay device, chemical reaction which causes coloration or discoloration by color reaction is performed after a predetermined period of time from when the liquid sample was added on the specimen, whereby determination based on visual observation is possible.
Measurement principle by immunochromatography is based on an assay method for identifying antigen or antibody utilizing the specificity of antigen-antibody reaction. The process is as follows: adding a liquid sample to an immunochromatographic specimen, a marker reagent dissolved by the liquid sample extends in infiltration direction and then a substance capable of being bound specifically to an analyte in the liquid sample is fixed, consequently coloration generates in a measurement range. Further, since the result of immunochromatographic analysis is determined by color reaction, it is recorded by the visual observation. In other words, since determination of the result of analysis is extremely easy, the range of use expands.
Such measurement by immunochromatography can be adopted for examinations of various analytes. In some cases with some analytes, however, only semi-quantitative result, at the most, is derived by this method when quantitative result is also required besides qualitative examination. Therefore, advent of any means to lead to more quantitative result of analysis has been desired.
To improve the above-mentioned problems, there is disclosed an immunochromatographic specimen in Japanese Published Patent Application Hei.8-240591. In an assaying method with this immunochromatographic specimen, after a sample was added to this specimen and reaction was done, a coloration degree detected on the specimen is measured as signals of absorption and reflection on the color-reacted portion, by using a detecting instrument. Further, Japanese Published Patent Application Hei.8-334511 discloses a quantitative assay method by immunochromatography using an immunochromatographic specimen, which is performed by taking a coloration degree given by analytic processing of a gradated image of the specimen previously converted after image-capturing by an image sensor.
However, in the case of the quantitative assay method using an immunochromatographic specimen as in the embodiments of the Japanese Published Patent Applications Hei.8-240591 and Hei.8-334511, the quantitative assay method is performed by adopting a suitable detecting instrument in accordance with the degree or level of coloration involved in color reaction. Conventionally, because an immunochromatographic specimen used in general can not be controlled artificially without mechanical control, infiltration rate of liquid sample is dominated by the permeability of the specimen. For example, owing to vaporization of liquid sample from side surfaces of the specimen, the amounts of analyte flowing out toward the measurement range sometimes become heterogeneity. Moreover, in each immunochromatographic specimen used for every measurement, the amount of liquid sample to be added and marker reagent to be added are not always fixed. Accordingly, there is a problem that these causes increase the dispersion of quantitative assay values that appear after a predetermined time.
Additionally, although an assay method by immunochromatography using immunochromatographic specimen is performed after a liquid sample is added and a predetermined period of time has passed, the marker reagent which permeates at this time can begin to dissolve simultaneously, and sometimes remains on the reaction layer as a background. If the value of this background is included in the coloration degree, the detecting instrument executes the measurement including such error. Therefore, quantitative measurement in short time with immunochromatographic specimen is hard to execute. Furthermore, residual amount of the marker reagent flowing out to the reaction layer is inconstant, and is different for every measurement or specimen to use, and thus obtained coloration degree at this time attributed in the error of the reading value greatly.
From the above-mentioned problem, performance of immunochromatographic quantitative assay was extremely close to semi-quantitative due to various standards for specimen and the dispersion of the added liquid sample, or the errors derived from background. Accordingly, the performance improvement of the specimen has been strongly required to enable more accurate measurement.
Further, when carrying out assay of a liquid sample including cell components such as whole blood or bacterial solution, using conventional immunochromatographic specimen, viscosity of the liquid sample and the presence of cell components cause clogging in a reaction layer carrier. Consequently, it takes much time for measurement, and desiccation of the liquid sample during measurement, if any, causes lack of reaction precision and quantitative efficiency. Thereby, simple and easy-handling immunochromatographic quantitative assay device has been strongly required for examination of whole blood, bacterial solution, or the like.
The present invention is made to solve the above-mentioned problems and has for its object to provide an immunochromatographic quantitative assay device and a manufacturing method thereof, which enables precise measurement without influenced by the character of the added liquid sample, accurate reading of coloration degree as a result of reaction of a liquid sample as an analyte and a marker reagent, and results in no influence in the residual amounts of the marker reagent by the background.
According to a first aspect of the present invention, there is provided a chromatographic quantitative assay device comprising: a non-moistenable reaction layer carrier support and a moistenable layer which is formed above the support and consists of one or arbitrary number of member portions, the moistenable layer comprising a portion for adding a liquid sample, a portion for holding a migratable marker reagent, a portion that performs binding reaction with a marker reagent which is provided with a substance capable of being bound specifically to an analyte in the liquid sample, and a portion for absorbing the liquid sample; and a liquid-impermeable sheet member which is provided being adhered to the upper surface and side surfaces except the portion for adding a liquid sample.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration.
According to a second aspect of the present invention, there is provided a chromatographic quantitative assay device comprising: a non-moistenable reaction layer carrier support and a moistenable layer which is formed above the support and consists of one or arbitrary number of member portions, the moistenable layer comprising a portion for adding a liquid sample, a portion for holding a constant quantity of migratable combination reagent which is previously prepared consisting of an analyte and a marker reagent, a portion that performs competitive binding reaction between an analyte in the liquid sample and the combination reagent provided with a substance capable of being bound specifically to the analyte in the liquid sample, and a portion for absorbing the liquid sample; and a liquid-impermeable sheet member which is provided being adhered to the upper surface and side surfaces except the portion for adding a liquid sample.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration.
According to a third aspect of the present invention, there is provided a chromatographic quantitative assay device comprising: a non-moistenable reaction layer carrier support and a moistenable layer which is formed above the support and consists of one or arbitrary number of member portions, the moistenable layer comprising a portion for adding a liquid sample where a substance for destroying cell components in the liquid sample is held, a portion for holding a migratable marker reagent, a portion that performs binding reaction with the marker reagent provided with a substance capable of being bound specifically to an analyte in the liquid sample, and a portion for absorbing the liquid sample; and a liquid-impermeable sheet member which is provided being adhered to the upper surface and side surfaces except the portion for adding a liquid sample.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration state of the liquid sample uniform by destroying cell components in the liquid sample, and to control the quantity of the sample and marker reagent which flow out in a constant time.
According to a fourth aspect of the present invention, there is provided a chromatographic quantitative assay device comprising: a non-moistenable reaction layer carrier support and a moistenable layer which is formed above the support and consists of one or arbitrary number of member portions, the moistenable layer comprising a portion for adding a liquid sample where a substance for destroying cell components in the liquid sample is held, a portion for holding a constant quantity of a migratable combination reagent which is previously prepared consisting of an analyte and a marker reagent, a portion that performs competitive binding reaction between an analyte in the liquid sample and the combination reagent provided with a substance capable of being bound specifically to the analyte in the liquid sample, and a portion for absorbing the liquid sample; and a liquid-impermeable sheet member which is provided being adhered to the upper surface and side surfaces except the portion for adding a liquid sample.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration state of the liquid sample uniform, and to control the quantity of the sample and combination reagent which flow out in a constant time.
According to a fifth aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member which is provided being adhered to the upper surface, or from the upper surface to the back surface, except the portion for adding a liquid sample.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, and to make the infiltration state of the liquid sample uniform. Furthermore, it is possible to make the marker reagent flow out at uniform concentration, and to restrain the affection of background values added on the values of color-reacted portions.
According to a sixth aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the reaction layer carrier support is removed; and the liquid-impermeable sheet member is provided on the periphery except the portion for adding a liquid sample.
Therefore, in this chromatographic quantitative assay device, it is possible to reduce the number of materials. Further, it is possible to make the infiltration state of the liquid sample uniform and to control the quantity of the sample and marker reagent which flow out in a constant time. Moreover, the liquid sample never leaks out of the gap, and the liquid sample never intrudes from the outside except a portion for adding the sample.
According to a seventh aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the vertical side surfaces of the liquid-impermeable sheet member is opened in the direction where the liquid sample infiltrates.
Therefore, in this chromatographic quantitative assay device, the air-releasing path at infiltration of liquid is restricted in only the infiltration direction, whereby it is possible that liquid sample and marker reagent flow out at uniform concentration.
According to an eighth aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the end of the liquid-impermeable sheet member is opened at an arbitrary position at the end of the chromatographic quantitative assay device in the direction where the liquid sample infiltrates.
Therefore, in this chromatographic quantitative assay device, the air-releasing path at infiltration of liquid is restricted in only the infiltration direction, whereby it is possible that liquid sample and marker reagent flow out at uniform concentration.
According to a ninth aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member is adhered by an adhesive made of an arbitrary adhesive.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample.
According to a tenth aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member is adhered by an arbitrary adhesive which hardens after adhesion.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample. Further, because the adhesive is hardened after adhesion, it is possible to prevent attachment of the paste to a cutter when the sensor is cut. Moreover, it is possible to facilitate mass production, as well as, to prevent transition of paste to reaction layer, thereby preventing transformation of reaction layer carrier or specific protein, whereby the performance of the chromatographic quantitative assay device can be maintained over a long period.
According to an eleventh aspect of the present invention, in a chromatographic quantitative assay device of the ninth aspect discussed above, the adhesive applied to the liquid-impermeable sheet member is made of thermoplastic resin.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the water, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample. Further, because the adhesive is hardened by cooling after adhesion, it is possible to prevent attachment of the paste to a cutter when the sensor is cut. Moreover, it is possible to facilitate mass production, as well as, to prevent transition of paste to reaction layer, thereby preventing transformation of reaction layer carrier or specific protein, whereby the performance of the chromatographic quantitative assay device can be maintained over a long period.
Here, the thermoplastic resin is a resin having the property that it is in a solid state at normal temperature, softens when heated to be processed, and solidifies when cooled thereafter. That is, it has the property that the plasticity is held reversibly by repetition of cooling and heating. The thermoplastic resin of this kind includes polyethylene, polypropylene, polychlorinated vinyl, polystyrene, polychlorinated vinylidene, fluororesin, methyl polymethacrylate, polyamide, polyester, polycarbonate, polyphenylene oxide, polyurethane, polyacetal, or the like.
According to a twelfth aspect of the present invention, in a chromatographic quantitative assay device of the ninth aspect discussed above, the adhesive applied to the liquid-impermeable sheet member is made of thermosetting resin.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample. Further, because the adhesive is hardened after adhesion and is not melted, it is possible to prevent attachment of the paste to a cutter when the sensor is cut. Moreover, it is possible to facilitate mass production, as well as, to prevent transition of paste to reaction layer, thereby preventing transformation of reaction layer carrier or specific protein, whereby the performance of a chromatographic quantitative assay device can be maintained over a long period.
Here, the thermosetting resin is a resin having the property that it softens when heated to be processed, but hardens by further heating and thus does not soften thereafter. The thermosetting resin of this kind includes urea resin, melamine resin, phenol resin, or the like.
According to a thirteenth aspect of the present invention, in a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member is made of an arbitrary transparent or semi-transparent liquid-impermeable material.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to enable visual detection when measuring the results on the detection region from the outside, whereby it is easy to perform the measurement.
According to fourteenth aspect of the present invention, a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member is made of an arbitrary semi-transparent or opaque liquid-impermeable material, and a transparent window is provided at the detection region.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to enable visual detection when measuring the results on the detection region from the outside, whereby it is easy to perform the measurement.
According to a fifteenth aspect of the present invention, in a chromatographic quantitative assay device of the third or fourth aspects discussed above, the substance for destroying cell components held at the portion for adding a liquid sample is a surfactant.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to avoid clogging to be caused by cell components in a liquid sample such as whole blood or bacterial solution, thereby to make the infiltration state uniform, resulting in more precise quantitative measurement in a short time.
According to a sixteenth aspect of the present invention, in a chromatographic quantitative assay device of the third or fourth aspects discussed above, the substance for destroying cell components held at the portion for adding a liquid sample is composed of chloride such as sodium chloride, potassium chloride.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to avoid clogging to be caused by cell components in a liquid sample such as whole blood or bacterial solution, thereby to make the infiltration state uniform, resulting in more precise quantitative measurement in a short time without inhibition of binding reaction.
According to a seventeenth aspect of the present invention, in a chromatographic quantitative assay device of the third or fourth aspect discussed above, the substance for destroying cell components held at the portion for adding a liquid sample is saponin series.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to avoid clogging to be caused by cell components in a liquid sample such as whole blood or bacterial solution, thereby to make the infiltration state uniform, resulting in more precise quantitative measurement in a short time.
According to an eighteenth aspect of the present invention, in a chromatographic quantitative assay device of the third or fourth aspects discussed above, the substance for destroying cell components held at the portion for adding a liquid sample is a substance having bacteriolytic effect such as lysozyme.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to avoid clogging to be caused by cell components in a liquid sample such as bacterial solution or thereby to make the infiltration state uniform, resulting in more precise quantitative measurement in a short time without inhibition of binding reaction.
According to a nineteenth aspect of the present invention, a manufacturing method of a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member, which is previously treated by performing aging at an arbitrary temperature, is adhered closely to the upper surface and side surfaces of the chromatographic quantitative assay device.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to accelerate the volatilization of organic solvent which brings adverse effects on the protein contained in the adhesive for the liquid-impermeable sheet member or on the reaction layer by performing aging, whereby preservation stability over a longer period is realized.
Here, the aging means leaving the liquid-impermeable sheet member for an arbitrary period of time under arbitrary and constant environment.
According to a twentieth aspect of the present invention, a manufacturing method of a chromatographic quantitative assay device of any of the first to fourth aspects discussed above, the liquid-impermeable sheet member is adhered closely to the upper surface and side surfaces of the chromatographic quantitative assay device by applying of an arbitrary pressure to the liquid-impermeable sheet member at an arbitrary temperature.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to adhere the liquid-impermeable sheet member onto the front surface and the side surfaces of the chromatographic quantitative assay device more solidly.
Here, the liquid-impermeable sheet member means e.g. an adhesive sheet material of heat seal type, therefore adhesion of a sheet material by laminating processing as seen in thermocompression bonding or the like is enabled.
According to a twenty-first aspect of the present invention, in a manufacturing method of a chromatographic quantitative assay device of the nineteenth or twentieth aspects discussed above, the adhesive applied to the liquid-impermeable sheet member has a characteristic of being melted and taking on stickiness at the temperature higher than normal temperatures and is hardened after adhesion.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample. Further, because the adhesive is hardened after adhesion, it is possible to prevent attachment of the paste to a cutter when the sensor is cut. Moreover, it is possible to facilitate mass production, as well as, to prevent transition of paste to reaction layer, thereby preventing transformation of reaction layer carrier or specific protein, whereby a manufacturing method of specimen that can keep the performance of the chromatographic quantitative assay device over a long period is provided.
Here, the adhesive is an adhesive having the property that it softens when heated and hardens when cooled after adhesion and, preferably, an adhesive having a low melting point as not to soften at normal temperature and to melt at lower temperature.
According to a twenty-second aspect of the present invention, a manufacturing method of a chromatographic quantitative assay device of the twenty-first aspect discussed above, the adhesive applied to the liquid-impermeable sheet member is made of thermoplastic resin.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample. Further, because the adhesive is hardened by cooling after adhesion, it is possible to prevent attachment of the paste to a cutter when the sensor is cut. Moreover, it is possible to facilitate mass production, as well as, to prevent transition of paste to reaction layer, thereby preventing transformation of reaction layer carrier or specific protein, whereby the performance of the chromatographic quantitative assay device can be maintained over a long period.
According to a twenty-third aspect of the present invention, a manufacturing method of a chromatographic quantitative assay device of the twenty-first aspect discussed above, the adhesive applied to the liquid-impermeable sheet member is made of thermosetting resin.
Therefore, in this chromatographic quantitative assay device, it is possible to prevent the vaporization of the moisture, to make the infiltration direction and infiltration state of the liquid sample uniform, and to make the liquid sample and marker reagent, which flow out in a constant time, flow out at uniform concentration, as well as, to prevent the liquid-impermeable sheet member from exfoliation by infiltration of the liquid sample. Further, because the adhesive is hardened after adhesion and is not melted, it is possible to prevent attachment of the paste to a cutter when the sensor is cut. Moreover, it is possible to facilitate mass production, as well as, to prevent transition of paste to reaction layer, thereby preventing transformation of reaction layer carrier or specific protein, whereby the performance of a chromatographic quantitative assay device can be maintained over a long period.